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A2547

Sigma-Aldrich

Monoclonal Anti-Actin, α-Smooth Muscle

clone 1A4, ascites fluid

Synonym(s):

Alpha Smooth Muscle Actin Antibody - Monoclonal Anti-Actin, α-Smooth Muscle, Alpha Smooth Muscle Actin Antibody Sigma, Anti-Alpha Smooth Muscle Actin Antibody, SMA

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About This Item

UNSPSC Code:
12352203
NACRES:
NA.41
conjugate:
unconjugated
application:
IF
IHC (f)
IHC (p)
WB
clone:
1A4, monoclonal
species reactivity:
human, mouse, rat, chicken, frog, canine, rabbit, guinea pig, goat, bovine, sheep, snake
citations:
2586
technique(s):
immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable using smooth muscle cells
immunohistochemistry (frozen sections): suitable using smooth muscle cells
indirect immunofluorescence: 1:400 using blood vessels in sections of human appendix
western blot: suitable using smooth muscle cells
Pricing and availability is not currently available.

biological source

mouse

Quality Level

conjugate

unconjugated

antibody form

ascites fluid

antibody product type

primary antibodies

clone

1A4, monoclonal

mol wt

antigen ~42 kDa

contains

15 mM sodium azide

species reactivity

human, mouse, rat, chicken, frog, canine, rabbit, guinea pig, goat, bovine, sheep, snake

technique(s)

immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable using smooth muscle cells
immunohistochemistry (frozen sections): suitable using smooth muscle cells
indirect immunofluorescence: 1:400 using blood vessels in sections of human appendix
western blot: suitable using smooth muscle cells

isotype

IgG2a

UniProt accession no.

application(s)

research pathology

shipped in

dry ice

storage temp.

−20°C

target post-translational modification

unmodified

Gene Information

human ... ACTA2(59)
mouse ... Acta2(11475)
rat ... Acta2(81633)

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General description

Actins are cytoskeletal proteins that regulate cell motility. Several scaffolding, signaling and actin-binding proteins regulate the interconversions between filamentous F-actins and monomeric G-actins. Cellular actins of various species have very similar immunological and physical properties. Hence, the use of monoclonal antibody to actin provides a specific and useful tool for studying the localization, structure and function of actin proteins. Actin 2 (ACTA2), the smooth muscle actin is encoded on human chromosome 10q23.31.
Mouse monoclonal anti-actin, α-smooth muscle antibody binds to actin from human, mouse, rat, bovine, chicken, frog, goat, guinea pig, rabbit, dog, sheep, and snake.
Monoclonal Anti-α Smooth Muscle Actin (mouse IgG2a isotype) is derived from the hybridoma produced by the fusion of mouse myeloma cells and splenocytes from an immunized mouse. The NH2 terminal synthetic decapeptide of α-smooth muscle actin coupled to keyhole limpet hemocyanin (KLH) was used as the immunogen. The isotype is determined using Sigma ImmunoTypeTM Kit (Sigma Stock No. ISO-1) and by a double diffusion immunoassay using Mouse Monoclonal Antibody Isotyping Reagents (Sigma Stock No. ISO-2).
The antibody (also known as anti-α-Sm-1) is specific for the single isoform of α-smooth muscle actin. It reacts specifically with α-smooth muscle actin in immunoblotting assays and labels smooth muscle cells in frozen or formalin-fixed, paraffin-embedded tissue sections.

Specificity

Monoclonal Anti-α Smooth Muscle Actin is specific for the single isoform α-actin. It reacts specifically with a smooth muscle actin using immunoblotting and indirect immunofluorescent labeling of formalin-fixed, paraffin embedded or frozen tissue sections. The antibody can be used for immunocytochemical localization of the α smooth muscle actin by means of immunoperoxidase, immunofluorescent, ELISA or immunoblot techniques. The antibody reacts with human, mouse, rat, bovine, chicken, frog, goat, guinea pig, rabbit, dog, sheep, and snake.

Immunogen

N-terminal synthetic decapeptide of α-smooth muscle actin.

Application

Immunocytochemistry was performed on smooth muscle cells from bovine aortas using the monoclonal anti-ACTA2 antibody. Cells were first grown on glass cover slips and fixed in 50% acetone/EtOH for 10 minutes at 4 degrees.
Mouse monoclonal anti-actin, α-smooth muscle antibody has been used for western blot assays using whole kidney homogenates in human anaplastic thyroid cancer cells. The product has also been used for immunohistochemical assays using mice tissue sections in liver sections and uterine sections.
Monoclonal Anti-Actin, α-Smooth Muscle antibody has been used in the detection of actin 2 in breast fibroblasts using immunofluorescence staining.
Paraffin embedded sections of rat testis tissue grafts were immunohistochemically stained with mouse monoclonal anti-smooth muscle actin.
IHC analysis of x-gal stained muouse cardiac tissue was performed using the primary antibody, mouse monoclonal anti-smooth muscle actin to identify myofibroblasts.

Biochem/physiol Actions

Actin is one of the most conserved eukaryotic proteins, it is expressed in mammals and birds as six isoforms characterized by electrophoresis and amino acid sequence analysis. Four of the six represent differentiation markers of muscle tissues. Actin isoforms show >90% overall sequence homology, but only 50-60% homology in their 18 NH2-terminal residues. The NH2-terminal region of actin appears to be a major antigenic region, and may be involved in the interaction of actin with other proteins such as myosin. It has been shown that the relative proportion of actin isoforms are different in smooth muscles of different organs and change within the same population of smooth muscle cells during development, pathological situations and different culture conditions.
Mutations in actin 2 is associated with dysfunction of smooth muscles in arteries and is implicated in coronary heart disease. Mutations in ACTA2 contributes to tear and enlargement of aorta in thoracic aortic aneurysms (TAAD).

Physical form

The antibody is provided as ascites fluid containing 15mM sodium azide as a preservative.

Storage and Stability

For continuous use, store at 2-8 °C for up to one month. For extended storage, solution may be frozen in working aliquots. Repeated freezing and thawing is not recommended. If slight turbidity occurs upon prolonged storage, clarify by centrifugation before use.

Other Notes

To view an Actin antibody selection guide, please visit www.sigmaaldrich.com/actin.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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Storage Class Code

10 - Combustible liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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Certificates of Analysis (COA)

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Natalie C Direkze et al.
Cancer research, 64(23), 8492-8495 (2004-12-03)
The role of myofibroblasts in tissue repair and fibrosis is well documented, but the source of these myofibroblasts is unclear. There is evidence of a circulating population of fibrocytes that can home to areas of injury and contribute to myofibroblast
Diverse functions of homologous actin isoforms are defined by their nucleotide, rather than their amino acid sequence.
Vedula P, et al.
eLife, 6, e31661-e31661 (2017)
Epithelial to mesenchymal transition in human breast cancer can provide a nonmalignant stroma.
Petersen OW, et al.
The American Journal of Pathology, 162(2), 391-402 (2003)
Transglutaminase-mediated cross-linking is involved in the stabilization of extracellular matrix in human liver fibrosis.
Grenard P, et al.
Journal of Hepatology, 35(3), 367-375 (2001)
Thoracic aortic aneurysm (TAAD)-causing mutation in actin affects formin regulation of polymerization.
Malloy LE, et al.
The Journal of Biological Chemistry, 287(34), 28398-28408 (2012)

Questions

1–8 of 8 Questions  
  1. Does this antibody work for IF staining of cultured cells on a cover slip? What secondary would you recommend?

    1 answer
    1. We do not test this product for IF on cells. However, it has been used in this application. You can use a secondary antibody like Product No. Anti-Mouse IgG (Fab specific)-FITC antibody produced in goat.

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  2. Which is the suggested concentration for IHC in FF tissue?

    1 answer
    1. The recommended concentration will be listed on the lot-specific Certificate of Analysis. For example, lot 0000221557 has an optimum concentration of 1:800.
      Please see the sample Certificate of Analysis for more information: https://www.sigmaaldrich.com/certificates/sapfs/PROD/sap/certificate_pdfs/COFA/Q14/A2547-.2ML0000246371.pdf

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  3. What is the Department of Transportation shipping information for this product?

    1 answer
    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

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  4. What is the molecular weight of actin, Products  A2066 and A2547?

    1 answer
    1. Actin has a molecular weight of 42 kDa.

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  5. Can Product A2547,  Monoclonal Anti-Actin, α-Smooth Muscle antibody detect beta actin?

    1 answer
    1. No, Product A2547,  Monoclonal Anti-Actin, α-Smooth Muscle antibody will not cross react with beta actin.

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  6. Is there a protocol for using Product A2547, Monoclonal Anti-Actin, α-Smooth Muscle antibody produced in mouse, in immunohistochemistry?

    1 answer
    1. This antibody is used in our Immunohistochemistry Kit, Product No. IMMH2.

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  7. What is the antibody concentration of Product A2547, Monoclonal Anti-Actin, α-Smooth Muscle antibody produced in mouse?

    1 answer
    1. This antibody is ascities fluid.  The amount of protein present and the amount of IgG1 present is indicated on the Lot-specific Certificate of Analysis.

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  8. Why do I have 3 bands on my western blot when using Product A2547, Monoclonal Anti-Actin, α-Smooth Muscle antibody produced in mouse?

    1 answer
    1. While actin should run as a 42 kDa band when detected by Western blotting, there are recent observations that cleavage products can be seen in cells undergoing apoptosis. Initial cleavage gives a band at 41 kDa, which may appear as a doublet on the blot. In addition, further cleavage results in a major band at 30 kDa and a small band at 14 kDa which have been shown to be cleavage products due to the action of interleukin 1 -converting enzyme. This may explain the detection of smaller bands when blotting with this antibody. This banding pattern may be detected with other actin antibodies as well. Kayalar, C., et al., Proc. Natl. Acad. Sci., USA, 93:2234-2238, (1996).

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